Sodium Chloride → 5'-Thymidylic acid

Participants

Reaction Steps

The thus prepared solution of the test compound was mixed with a solution containing 50 mM of Tris-HCl buffer (pH 8.3), 8 mM of MgCl2, 30 mM of NaCl, 50 mM of dithiothreitol (made by Wako Pure Chemical Industries Ltd.), 0.2 mM of thymidine-5'-triphosphate (made by Pharmacia K. K.) and 6 U/ml of reverse transcriptase derived from M-MLV (made by Pharmacia K. K.), and preincubated at 37° C. for 30 minutes. After there was added thereto 10 μg/ml of polyadenylic acid (made by PL Biochemicals Co., Ltd.), 0.01 U/ml of oligodeoxy thymidylic acid (made by Pharmacia K. K.) and 10 μCi/ml of [methyl-3H] thymidine-5'-triphosphate (made by Amersham Japan Co., Ltd., 47 Ci/mmol) to give a reaction mixture, the mixture was further incubated at 37° C. for 30 minutes, followed by cooling with ice to stop the reaction.

Provenance & Evidence